Synthesis of ionizable lipopolymers using split-Ugi reaction for pulmonary delivery of various size RNAs and gene editing
Kseniya Yu. Vlasova, Alexander M. Kerr, Nathan D. Pennock, Antony Jozić, Deepak Kumar Sahel, Milan Gautam, Namratha Turuvekere Vittala Murthy, Amanda J. Roberts, Muhammad Waqar Ali, Kelvin D. MacDonald, Sue Walker, Robert Luxenhofer, Gaurav Sahay
Abstract
We present an efficient method for synthesizing cationic poly(ethylene imine) derivatives using the multicomponent split-Ugi reaction to create a library of functional ionizable lipopolymers. Here we show 155 polymers, formulated into polyplexes, to establish structure-activity relationships essential for endosomal escape and transfection. A lead structure is identified, and lipopolymer-lipid hybrid nanoparticles are developed to deliver mRNA to lung endothelium and immune cells, including T cells, with low in vivo toxicity. These nanoparticles show significant improvements in mRNA delivery to the lung compared to in vivo-JetPEI® and demonstrate effective delivery of therapeutic mRNA(s) of various sizes. IL-12 mRNA-loaded nanoparticles delay Lewis Lung cancer progression, while human CFTR mRNA restores CFTR protein function in CFTR knockout mice. Additionally, we demonstrate in vivo CRISPR-Cas9 mRNA delivery, achieving gene editing in lung tissue and successful PD-1 knockout in T cells in mice. These results highlight the platform’s potential for systemic gene therapy delivery. Development of safe and effective non-viral delivery systems is important for broadening clinical applications of mRNA delivery. Here, Vlasova et al. create a library of ionizable lipopolymers using split-Ugi reaction to develop nanoparticles for targeted pulmonary delivery of mRNA.